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Transposon Mutagenesis in a Pathogenic Strain of Listeria monocytogenes

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Title: Transposon Mutagenesis in a Pathogenic Strain of Listeria monocytogenes
Authors: Nitta, Sharon
Issue Date: 15 Jan 2014
Publisher: University of Hawaii at Manoa
Abstract: Listeria monocytogenes is a ubiquitous bacterium which has recently been implicated in several food-related epidemics of serious disease (listeriosis). The monoclonal antibody MAb 198 has revealed the presence of a prominent temperature-regulated antigen on the surface of Listeria monocytogenes. We have pursued the characterization of the antigen and its possible involvement in bacterial growth and virulence. Transposon Tn916 was inserted into Listeria monocytogenes' DNA to induce a mutation in the genetic determinant needed for antibody recognition of MAb 198 by the antigen. The transposon was transferred by conjugation of a streptomycin-resistant Listeria monocytogenes recipient strain with Enterococcus faecalis RH110 on nitrocellulose filters. Among the streptomycin-resistant transconjugants, nonmotile mutants were detected using 0.4% trypticase soy agar (TSA) plates. Mutants lacking MAb 198 activity were not easily detected using immunoblotting and enzyme-linked immunoassay (ELISA) techniques. Several possible mutants may have been isolated, but further testing needs to be done to confirm their identity.
Pages/Duration: ii, 37 pages
Rights: All UHM Honors Projects are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission from the copyright owner.
Appears in Collections:Honors Projects for Biology

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